Objective:To observe the effect of Pidotimod and Thalidomid to enhance the immune response and protective immunity induced by the epitope Vaccines From W2b2a of Toxoplasma gondii in mice.Methods:Mice were immunized intramuscularly with Pidotimod and pcDNA3-W2b2a,thalidomide and pcDNA3-W2b2a,respectively,then the immune response and the survival time of mouse attacked by Toxoplasma gondii were observed.Results:After the immunization,the level of IFN-γin sera of mice inculated with pcDNA3-W2b2a and Pidotimod,pcDNA3-W2b2a and Freund adjuvant,pcDNA3-W2b2a and Thalidomid were significantly higher than pcDNA3-W2b2a (all P<0.05).After the immunization,IgG,CD4+/CD8+T cell ratio ,proliferation of T cell induced by pcDNA3-W2b2a and Pidotimod,pcDNA3-W2b2a and Freund adjuvant were higher than pcDNA3-W2b2a,pcDNA3-W2b2a and thalidomid ( all P<0.05).After challenged with highly virulent tachyzoites,the mean survival time in immunized groups were significantly longer than control group (all P<0.05).Conclusion:Pidotimod ,Thalidomid adjuvant can increase protective immunity of epitope Vaccines From W2b2a of Toxoplasma gondii in mice.

OBJECTIVETo introduce the experience of treating nonunions of humeral fractures with interlocking intramedullary nailing.

METHODSTwelve patients with humeral nonunions were treated with interlocking intramedullary nailing. The time interval between trauma and surgery was 10.5 months on average. Open reduction with anterograde approach was performed. Axial compression was specially applied to the fracture site with humeral nail holder after insertion of distal locked screws. Iliac bone grafting was added.

RESULTSThe average follow-up period was 21 months (ranging 9-51 months). All patients achieved osseous union 5.8 months after treatment on average. Eleven patients had good functions of the shoulder joints and the upper extremities. No patient experienced any permanent neurological deficit. Refracture of the original ununited region occurred in one patient after removal of the internal fixator one year later, but union was achieved after closed re-intramedullary nailing fixation.

CONCLUSIONHumeral interlocking intramedullary nailing is an effective alternative treatment for humeral nonunion.

Adult ; Aged ; Bone Nails ; Bone Transplantation ; methods ; Female ; Fracture Fixation, Intramedullary ; instrumentation ; Fractures, Ununited ; surgery ; Humans ; Humeral Fractures ; diagnostic imaging ; surgery ; Ilium ; transplantation ; Male ; Middle Aged ; Radiography ; Treatment Outcome

To investigate the effects of telmisartan on steatohepatitis (NASH) in rats by activating peroxisome proliferator-activated receptor gamma (r). Thirty male SD rats were randomized into normal control group, NASH control group and telmisartan prevention group. Normal control group was given standard food and the other two groups were given high fat diet for 16 weeks to induce NASH. Prevention group was given telmisartan (5 mg.kg-1.d-1) for 4 weeks by intragastric adminstration after 12 weeks. At the end of the 16th week, all the rats were sacrificed. Pathological changes of liver were observed by optical microscopy. Serum alanine aminotransferase(ALT), aspartate aminotransferase(AST), fasting blood glucose(FBG), fasting insulin(FINS), HOMA-IR(homeostasis model assessment insulin resistance), Serum TNF-a and adiponectin were detected and analyzed.Western blot and RT-PCR were used to detect PPARr expression in hepatic tissues on protein and mRNA levels. (1) Rats were successfully modeled. The liver tissue samples were divided into 4 degrees (F0 - 4) based on total fatty degeneration of liver cells.There was one rat reached F3 and nine rats reached F4 in NASH group, one rat reached F1, six rats reached F2 and three rats reached F3 in prevention group. Inflammatory activity scores of hepatic tissues in the model group were 2.67+/-0.25, while that in the control group was 0 (U=15 and P is less than to 0.01), in the prevention group were 2.67+/-0.25 and 1.36+/-0.12 (U=24 and P is less than to 0.05 ). (2) The levels of serum ALT, AST, FBG, FINS, TNFa and HOMA-IR in the model group were increased than those in the control group( the vaules of q were 13.130, 6.472, 6.909, 26.619, 14.591 and 49.683 respectively, P less than 0.01). The levels of serum ALT, FINS, FBG, TNFa and HOMA-IR in the prevention group were decreased as compared to the model group (the vaules of q were 7.024, 4.145, 14.829, 13.195 and 31.991 respectively, P less than 0.01 ). (3) The serum adiponectin, PPARrmRNA and protein in liver tissues of the model group were lower than those in the control group (q values were 10.696, 8.679 and 16.762 respectively, P is less than to 0.05).The data in the prevention group were higher as compared to the model group(q values were 3.879,3.079,6.400, P is less than to 0.05 respectively). HOMA-IR was positively correlated with the expression of TNFa but negatively correlated with the expression of adiponectin (r = 0.927, P is less than to 0.01; r = -0.891, P is less than to 0.01, respectively). Telmisartan may has preventive effect on rats with steatohepatitis (NASH) by a mechanism of activating peroxisome proliferator-activated receptor r.
Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Fatty Liver ; Insulin Resistance ; Non-alcoholic Fatty Liver Disease ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo construct a reasonable substitute for the autograft bone in vitro and transplant it back into the rabbit models to induce the spine fusion.

METHODSThe bone marrow stem cell from the seven New Zealand rabbits were cultured. Recombinant human bone morphogenetic protein-4 (rhBMP-4) that has been proved to be bioactive was obtained by the way of genetic engineering. Using the vacuum freezing machine to mix a certain quantity of rhBMP-4 into type I collagen to form a new kind of carrier. Animal model of spine facet process fusion was used. Bone marrow stem cells combined with rhBMP-4 and I type collagen were implanted between the facet process to induce the spine fusion. type I collagen and bone marrow stem cell was used in the controlled group.

RESULTSNew bone formation was obvious in the test group. The facet joint was fused very well in this side. No bone formation was present on the other side.

CONCLUSIONSThe new composite: bone marrow stem cells, rhBMP-4 and type I collagen was an ideal kind of substitute for the autograft bone.

Animals ; Bone Marrow Cells ; cytology ; Bone Morphogenetic Protein 4 ; Bone Morphogenetic Proteins ; biosynthesis ; genetics ; Bone Substitutes ; therapeutic use ; Bone Transplantation ; Cells, Cultured ; Collagen Type I ; chemistry ; Implants, Experimental ; Rabbits ; Recombinant Proteins ; biosynthesis ; genetics ; Spinal Fusion ; methods ; Stem Cell Transplantation ; Stromal Cells ; cytology ; Tissue Engineering

BACKGROUNDRecent studies have demonstrated that dexamethasone (DEX) interferes with immune responses by targeting key functions of dendritic cells (DCs) at the earliest stage. However, the cellular and molecular mechanisms are still incompletely understood. This study aimed to explore the possible mechanisms by investigating the roles of DEX on differentiation, maturation & function of murine DCs and the effects of DEX on DCs via Toll-like receptor 4 (TLR4)-nuclear factor (NF)-kappaB mediated signal pathway.

METHODSImmature DCs (imDCs) were cultured from murine bone marrow (BM) cells. We added DEX into culture medium at different time. The expression of CD11c, CD86 and I-A(b) (mouse MHC class II molecule) was determined by flow cytometry. We determined the expression of NF-kappaB and its inhibitory protein I-kappaBalpha by electrophoretic mobility shift assay (EMSA) and Western blotting, respectively. The productions of interleukin (IL)-12p70 and IL-10 in cell culture supernatants were determined by enzyme-linked immunosorbent assay (ELISA).

RESULTSDEX impaired differentiation of DCs from murine bone marrow progenitors, and inhibited lipopolysaccharide (LPS) induced maturation of DCs. DEX significantly inhibited NF-kappaB expression of normal DCs, the higher the DEX concentration or the longer the DEX treatment time, the more obvious the effect. However, DEX had little effect on LPS-induced NF-kappaB activation, and partially impaired LPS-induced I-kappaBalpha degradation. DEX significantly decreased LPS induced IL-12p70 production by DCs. Interestingly, our results showed a synergistic effect between DEX and LPS on the production of IL-10 by DCs.

CONCLUSIONSDEX inhibits the differentiation and maturation of murine DCs involved in TLR4-I-kappaB-NF-kappaB pathway, and also indirectly impairs Th1 development and interferes with the Th1-Th2 balance through IL-12 and/or IL-10 secretion by DCs.

Animals ; Blotting, Western ; Bone Marrow Cells ; cytology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Dendritic Cells ; cytology ; metabolism ; Dexamethasone ; pharmacology ; Electrophoretic Mobility Shift Assay ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Male ; Mice ; NF-kappa B ; metabolism ; Signal Transduction ; drug effects ; Toll-Like Receptor 4 ; metabolism

Objective To study the mechanism of tumor necrosis factor-related apoptosis-inducing ligand(TRAIL)-induced apoptosis of glioma U87 cells.Methods Human glioma U87 cells were treated with human recombinant soluble TRAIL(rsTRAIL),and the cell apoptosis was detecmd with flow cytometry with AnnexinV-FITC/PI double staining.Flow cytometry with DiOC6 staining was used to assess the changes in mitochondrial transmembrane potential(△ψm).The relative activity of caspase-3,-8 and-9 Was measmed by colorimetric assay,and the concentration of cytoplasmic cytochrome C(cyt C) determined using enzyme-linked immunosorbem assay.The effects ofcaspase-8 inhibitor(Z-IETD-fmk)on rsTRAIL-induced apoptosis,△ψm,caspase-3,-8 and-9 activities and cyt C concentration were observed. Results RsTRAIL tinle-dependently induced apoptosis and progressive collapse of △ψm in glioma U87 cells,resulting also in caspase-3,-8 and-9activation and elevated cytC concentration.Caspase-8 inhibitor partially antagonized these biological effects induced by rsTRAIL in U87 cells.Conclusion TRAIL initiates a cascade of mitochondrial events by activating caspase-8 and induces apoptosis of glioma U87 cells.

Anticarcinogenic Agents ; administration & dosage ; isolation & purification ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Non-Small-Cell Lung ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Humans ; Isothiocyanates ; administration & dosage ; isolation & purification ; pharmacology ; Lung Neoplasms ; pathology ; Plants, Medicinal ; chemistry ; Protein Binding ; Raphanus ; chemistry ; Tubulin ; chemistry ; metabolism

AIM:To investigate the effects of xeroderma pigmentosum group D ( XPD) gene on the prolifera-tion of human umbilical arterial smooth muscle cells ( HUASMCs) induced by oxidized low-density lipoprotein ( Ox-LDL) . METHODS:The recombinant plasmid pEGFP-N2/XPD was transfected into HUASMCs by liposome .The cells were di-vided into blank control group , pEGFP-N2 group, pEGFP-N2/XPD group, Ox-LDL group, Ox-LDL+pEGFP-N2 group and Ox-LDL+pEGFP-N2/XPD group.The proliferation rate of the cells was detected by MTT and EdU assays .The apop-totic rate and cell cycle distribution were analyzed by flow cytometry .The protein levels of XPD, caspase-3, Bcl-2 and Bax were determined by Western blot .RESULTS:Compared with blank control group , the expression of XPD was increased in pEGFP-N2/XPD group (P<0.05).According to the results of MTT and EdU assays , the cell proliferation in pEGFP-N2/XPD group was reduced compared with blank control group (P<0.05).Compared with Ox-LDL group, the cell prolifera-tion in Ox-LDL+pEGFP-N2/XPD group was significantly inhibited (P<0.05).According to the results of flow cytome-try, the cell proportion of S phase decreased and the G 0/G1-phase cell proportion increased significantly in pEGFP-N2/XPD group and Ox-LDL+pEGFP-N2/XPD group compared with blank control group and Ox-LDL group, repectively (P<0.05).Compared with blank control group and Ox-LDL group, the protein level of Bcl-2 decreased and the protein levels of Bax and cleaved caspase-3 increased in pEGFP-N2/XPD group and Ox-LDL +pEGFP-N2/XPD group, respectively (P<0.05).CONCLUSION:XPD inhibits the proliferation of HUASMCs and promotes their apoptosis , and reduces the promoting effect of Ox-LDL on the proliferation of HUVSMCs .XPD may be the target for treatment of atherosclerosis .

OBJECTIVETo culture and study the osteogenic characteristics of human bone marrow-derived mesenchymal stem cells (hBMMSCs).

METHODShBMMSCs were separated and cultured from human iliac crest marrow. Growth kinetics of hBMMSCs was studied by growth curve. Under the osteoinductive culture, osteogenic differentiation of hBMMSCs was tested by alkaline phosphatase (ALP). Osteogenic functions of hBMMSCs in vitro and in vivo were also respectively detected by von Kossa stain and by transplanting hydroxyapatite/tricalcium phosphate ceramics (HA/TCP) with hBMMSCs.

RESULTShBMMSCs were cultured successfully. The growth curve of the second passage of BMMSCs indicated that the time of population doublings was about 3.5 days. The results of ALP stain were evident by the significant increase in ALP activity after hBMMSCs cultured in osteoinductive medium. Some mineralized nodules were detected by von Kossa stain at nineteenth day of osteoinductive culture. In vivo assay, histological evalution showed bone formation in 3 months after grafts of HA/TCP with hBMMSCs.

CONCLUSIONSOsteoinductive solution can induce hBMMSCs to differentiate osteogenetic cell lines. Mineralized nodules and bone formation were found in vitro and in vivo assay. The results demonstrate that hBMMSCs have the potential for osteogenesis.

Adolescent ; Adult ; Alkaline Phosphatase ; analysis ; Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; Child ; Female ; Humans ; Male ; Mesenchymal Stromal Cells ; cytology ; Mice ; Osteogenesis

OBJECTIVETo investigate the clinical characteristics, treatment options and causes of misdiagnosis of ipsilateral femoral neck and shaft fractures.

METHODSAmong 20 patients with ipsilateral femoral neck and shaft fractures, 19 were treated operatively and 1 was treated conservatively. Sixteen cases of femoral shaft fractures were treated by open reduction and internal fixation with compressive plate, and 2 cases were treated with interlocking intramedullary nailing. Eighteen femoral neck fractures were treated with cannulated screws. Another patient was treated with proximal femoral nail to fix both the neck and shaft. Delayed diagnosis for femoral neck fractures occurred in 2 cases preoperatively.

RESULTSA total of 19 patients were followed up. The follow up period ranged from 5 to 48 months with an average of 15 months. All the fractures were healed.

CONCLUSIONFor case of femoral shaft fracture caused by high energy injury, an AP pelvic film should be routinely taken. Once the femoral neck fracture is recognized, operative reduction and fixation should be performed in time. Femoral neck and shaft fractures should be fixed separately.

Adult ; Aged ; Female ; Femoral Fractures ; diagnosis ; surgery ; Femoral Neck Fractures ; diagnosis ; surgery ; Fracture Fixation, Internal ; Humans ; Male ; Middle Aged